ArrestinDb

grm1_human

NAME

4OR2

ORGANISM

Human (Homo sapiens)

PROTEIN NAME

grm1_human

SOURCE

SWISSPROT

SEQUENCE

Perspective: Wild-type Construct


N-term
M	V	G	L	L	L	F	F	F	P
10



A	I	F	L	E	V	S	L	L	P
20



R	S	P	G	R	K	V	L	L	A
30



G	A	S	S	Q	R	S	V	A	R
40



M	D	G	D	V	I	I	G	A	L
50



F	S	V	H	H	Q	P	P	A	E
60



K	V	P	E	R	K	C	G	E	I
70



R	E	Q	Y	G	I	Q	R	V	E
80



A	M	F	H	T	L	D	K	I	N
90



A	D	P	V	L	L	P	N	I	T
100



L	G	S	E	I	R	D	S	C	W
110



H	S	S	V	A	L	E	Q	S	I
120



E	F	I	R	D	S	L	I	S	I
130



R	D	E	K	D	G	I	N	R	C
140



L	P	D	G	Q	S	L	P	P	G
150



R	T	K	K	P	I	A	G	V	I
160



G	P	G	S	S	S	V	A	I	Q
170



V	Q	N	L	L	Q	L	F	D	I
180



P	Q	I	A	Y	S	A	T	S	I
190



D	L	S	D	K	T	L	Y	K	Y
200



F	L	R	V	V	P	S	D	T	L
210



Q	A	R	A	M	L	D	I	V	K
220



R	Y	N	W	T	Y	V	S	A	V
230



H	T	E	G	N	Y	G	E	S	G
240



M	D	A	F	K	E	L	A	A	Q
250



E	G	L	C	I	A	H	S	D	K
260



I	Y	S	N	A	G	E	K	S	F
270



D	R	L	L	R	K	L	R	E	R
280



L	P	K	A	R	V	V	V	C	F
290



C	E	G	M	T	V	R	G	L	L
300



S	A	M	R	R	L	G	V	V	G
310



E	F	S	L	I	G	S	D	G	W
320



A	D	R	D	E	V	I	E	G	Y
330



E	V	E	A	N	G	G	I	T	I
340



K	L	Q	S	P	E	V	R	S	F
350



D	D	Y	F	L	K	L	R	L	D
360



T	N	T	R	N	P	W	F	P	E
370



F	W	Q	H	R	F	Q	C	R	L
380



P	G	H	L	L	E	N	P	N	F
390



K	R	I	C	T	G	N	E	S	L
400



E	E	N	Y	V	Q	D	S	K	M
410



G	F	V	I	N	A	I	Y	A	M
420



A	H	G	L	Q	N	M	H	H	A
430



L	C	P	G	H	V	G	L	C	D
440



A	M	K	P	I	D	G	S	K	L
450



L	D	F	L	I	K	S	S	F	I
460



G	V	S	G	E	E	V	W	F	D
470



E	K	G	D	A	P	G	R	Y	D
480



I	M	N	L	Q	Y	T	E	A	N
490



R	Y	D	Y	V	H	V	G	T	W
500



H	E	G	V	L	N	I	D	D	Y
510



K	I	Q	M	N	K	S	G	V	V
520



R	S	V	C	S	E	P	C	L	K
530



G	Q	I	K	V	I	R	K	G	E
540



V	S	C	C	W	I	C	T	A	C
550



K	E	N	E	Y	V	Q	D	E	F
560



T	C	K	A	C	D	L	G	W	W
570



P	N	A	D	L	T	G	C	E	P
580



I	P	V	R	Y	L	E	W	S	N
590


TM1
I	E	S	I	I	A	I	A	F	S
600



C	L	G	I	L	V	T	L	F	V
610



T	L	I	F	V	L	Y	R	D	T
620


ICL1						TM2
P	V	V	K	S	S	S	R	E	L
630



C	Y	I	I	L	A	G	I	F	L
640



G	Y	V	C	P	F	T	L	I	A
650


ECL1				TM3
K	P	T	T	T	S	C	Y	L	Q
660



R	L	L	V	G	L	S	S	A	M
670



C	Y	S	A	L	V	T	K	T	N
680


						ICL2
R	I	A	R	I	L	A	G	S	K
690



K	K	I	C	T	R	K	P	R	F
700


	TM4
M	S	A	W	A	Q	V	I	I	A
710



S	I	L	I	S	V	Q	L	T	L
720



V	V	T	L	I	I	M	E	P	P
730


ECL2
M	P	I	L	S	Y	P	S	I	K
740



E	V	Y	L	I	C	N	T	S	N
750


TM5
L	G	V	V	A	P	L	G	Y	N
760



G	L	L	I	M	S	C	T	Y	Y
770


				ICL3
A	F	K	T	R	N	V	P	A	N
780


		TM6
F	N	E	A	K	Y	I	A	F	T
790



M	Y	T	T	C	I	I	W	L	A
800



F	V	P	I	Y	F	G	S	N	Y
810


TM7
K	I	I	T	T	C	F	A	V	S
820



L	S	V	T	V	A	L	G	C	M
830



F	T	P	K	M	Y	I	I	I	A
840


	H8
K	P	E	R	N	V	R	S	A	F
850


			C-term
T	T	S	D	V	V	R	M	H	V
860



G	D	G	K	L	P	C	R	S	N
870



T	F	L	N	I	F	R	R	K	K
880



A	G	A	G	N	A	N	S	N	G
890



K	S	V	S	W	S	E	P	G	G
900



G	Q	V	P	K	G	Q	H	M	W
910



H	R	L	S	V	H	V	K	T	N
920



E	T	A	C	N	Q	T	A	V	I
930



K	P	L	T	K	S	Y	Q	G	S
940



G	K	S	L	T	F	S	D	T	S
950



T	K	T	L	Y	N	V	E	E	E
960



E	D	A	Q	P	I	R	F	S	P
970



P	G	S	P	S	M	V	V	H	R
980



R	V	P	S	A	A	T	T	P	P
990



L	P	S	H	L	T	A	E	E	T
1000



P	L	F	L	A	E	P	A	L	P
1010



K	G	L	P	P	P	L	Q	Q	Q
1020



Q	Q	P	P	P	Q	Q	K	S	L
1030



M	D	Q	L	Q	G	V	V	S	N
1040



F	S	T	A	I	P	D	F	H	A
1050



V	L	A	G	P	G	G	P	G	N
1060



G	L	R	S	L	Y	P	P	P	P
1070



P	P	Q	H	L	Q	M	L	P	L
1080



Q	L	S	T	F	G	E	E	L	V
1090



S	P	P	A	D	D	D	D	D	S
1100



E	R	F	K	L	L	Q	E	Y	V
1110



Y	E	H	E	R	E	G	N	T	E
1120



E	D	E	L	E	E	E	E	E	D
1130



L	Q	A	A	S	K	L	T	P	D
1140



D	S	P	A	L	T	P	P	S	P
1150



F	R	D	S	V	A	S	G	S	S
1160



V	P	S	S	P	V	S	E	S	V
1170



L	C	T	P	P	N	V	S	Y	A
1180



S	V	I	L	R	D	Y	K	Q	S
1190



S	S	T	L


hemagglutin


FLAG


His(10)


TEV proteas


BRIL (b562R


	N-term
	I	P	V	R	Y	L	E	W	S
589


	TM1
N	I	E	S	I	I	A	I	A	F
599



S	C	L	G	I	L	V	T	L	F
609



V	T	L	I	F	V	L	Y	R	D
619


ICL1							TM2
T	P	V	V	K	S	S	S	R	E
629



L	C	Y	I	I	L	A	G	I	F
639



L	G	Y	V	C	P	F	T	L	I
649


	ECL1				TM3
A	K	P	T	T	T	S	C	Y	L
659



Q	R	L	L	V	G	L	S	S	A
669



M	C	Y	S	A	L	V	T	K	T
679



N	R	I	A	R	I	L	A	G	S
689


ICL2
K	K	K	I	C	T	R	K	P	R
699


		TM4
F	M	S	A	W	A	Q	V	I	I
709



A	S	I	L	I	S	V	Q	L	T
719



L	V	V	T	L	I	I	M	E	P
729


ECL2
P	M	P	I	L	S	Y	P	S	I
739



K	E	V	Y	L	I	C	N	T	S
749


TM5
N	L	G	V	V	A	P	L	G	Y
759



N	G	L	L	I	M	S	C	T	Y
769


					ICL3
Y	A	F	K	T	R	N	V	P	A
779


			TM6
N	F	N	E	A	K	Y	I	A	F
789



T	M	Y	T	T	C	I	I	W	L
799



A	F	V	P	I	Y	F	G	S	N
809


TM7
Y	K	I	I	T	T	C	F	A	V
819



S	L	S	V	T	V	A	L	G	C
829



M	F	T	P	K	M	Y	I	I	I
839


		H8
A	K	P	E	R	N	V	R	S	A
849


				C-term
F	T	T	S	D	V	V	R	M	H
859



V

SCHEMATIC

Perspective: Wild-type Construct

N-term

TM1

ICL1

TM2

ECL1

TM3

ICL2

TM4

ECL2

TM5

ICL3

TM6

ECL3

TM7

H8

C-term

hemagg

FLAG

His(10

TEV pr

BRIL (

N-term

TM1

ICL1

TM2

ECL1

TM3

ICL2

TM4

ECL2

TM5

ICL3

TM6

ECL3

TM7

H8

C-term

DIAGRAMS

MUTATIONS

Position	Wild-type	Mutated

DELETIONS

from 1 to 580

from 861 to 1194

MODIFICATIONS

AUXILIARY PROTEINS

Protein_type: signal

Name: hemagglutinin

Sequence: None

Deletions:

Positions: N-term_0 None:None

Presence in Crystal:NO

Remarks: None

Protein_type: tag

Name: FLAG

Sequence: None

Deletions:

Positions: N-term_1 None:None

Presence in Crystal:NO

Remarks: None

Protein_type: tag

Name: His(10)

Sequence: None

Deletions:

Positions: N-term_2 None:None

Presence in Crystal:NO

Remarks: None

Protein_type: prot_cleavage

Name: TEV protease

Sequence: None

Deletions:

Positions: N-term_3 None:None

Presence in Crystal:NO

Remarks: None

Protein_type: fusion

Name: BRIL (b562RIL)

Sequence: None

Deletions:

Positions: N-term_4 None:None

Presence in Crystal:YES

Remarks: None

EXPRESSION

Expression method	Host cell type	Host cell	Remarks
Baculovirus	Sf9	Insect	Bac - to - Bac Baculovirus Expression System (Invitrogen)

SOLUBILIZATION

Solvent type	Item	Concentration
additive	CHS	0.2 %w/v
detergent	DDM	1 %w/v

Remarks:

PURIFICATION

Purification type	Description
Alkylation by Iodoacetamide	None
Proteolysis by TEV protease	None

Remarks:None

CRYSTALLIZATION

METHOD: in meso/LCP

TYPE: lipidic cubic phase (LCP) (monoolein (9.9 MAG))

PROTEIN CONCENTRATION: 50-80 mg/ml

Temperature: 20°C

ph : 7.0-7.0

Remarks: molten lipid constituted by 90% (w/w) of monoolein and 10% (w/w) of cholesterol

Chemical lists

Type	Item	Concentration
Additional
unknown	HEPES	100 mM
unknown	PEG400	27-30% v/v
unknown	(NH4)2HPO4	80-120 mM
unknown	TCEP	4-8 mM
Detergent
detergent	DDM	0.05 w/v
Lipid
lipid	CHS	0.01 %w/v